alpha-D-mannosidase,p-nitrophenyl-alpha-mannosidase,alpha-D-mannopyranosidase,alphamannosidase,exo-alpha-mannosidase

AlphaMannosidasefromJackBeancleavesα(1-2,3,6)-linkedmannose.

ThisenzymeisoftenusedinconjunctionwithCoreAlpha-(1-6)Mannosidase(E-AM02)ifanoncleavablecoreα(1-6)mannoseispresentonthesubstrate.

SourceJackBean

EC3.2.1.24

Contents
Alpha-(1-2,3,6)Mannosidasein150mMsodiumphosphate,0.1mMZnCl2pH7.5.(pH7.5).

Includedwith20µLand60µLpacksizes:
200µl5xReactionBuffer5.0(250mMsodiumphosphate,pH5.0).

SpecificActivity>3U/mg
Activity>10U/ml

Molecularweighttwopolypeptidesof64,000and44,000daltons

Suggestedusage
1.Addupto1nmolofoligosaccharide.
2.Adddeionizedwaterto15µl.
3.Add4µl5xReactionBuffer5.0.
4.Add1µLofenzyme
5.Incubatetenminutesat37˚C.

SpecifictyAllAlpha-(1-2,3,6)-linkedmannose

SpecificActivityAssay
OneunitofAlphaMannosidaseisdefinedastheamountofenzymerequiredtohydrolyze1µmoleofp-nitrophenyl-alpha-p-mannosidetop-nitrophenolin1minuteatpH5.0and37˚C.

StorageStoreenzymeat4˚C.

PurityEachlotofα(1-2,3,6)mannosidaseistestedforcontaminatingsubstancesbyincubatingtheenzymefor24hoursat37°Cwithsubstratesindicatedinthetablebelow.Nodetectableactivityisevidentforanyofthesepotentialcontaminants.Thedetectionlimitofthisassayis5μU/mL(IUB).

Fortheproteaseassay,10μgofdenaturedBSAisincubatedfor24hourswith2μlofenzyme.AnalysisoftheBSAbandafterSDS-PAGEshouldshownoevidenceofdegradation.

StABIlityStableatleast12monthswhenstoredproperly.Severaldaysexposuretoambienttemperatureswillnotreduceactivity.

AlphaMannosidaseReferences

Snaith,S.M.,andG.A.Levvy.PurificationandPropertiesofα-D-MannosidasefromJack-Beanmeal.Biochem.J.110:633-670(1968)

GaneshKumar,B.S.,Pohlrntz,G.,Shculte,M.,Mormann,M.SivaKumar,N.Jackbeanα-mannosidase:aminoacidsequencingandN-glycosylationanalysisofavaluableglycomicstool.GlycoBIOLOGy24(3):252-61(2014)